JPH10182613A - Tetraphydroisoquinoline derivative - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new compound useful for preventing and treating autoimmune disease, having inhibitory action on dipepetidyl peptidase IV, capable of suppressing T cells. SOLUTION: This new compound is shown by formula I [R<1> is an amino- protecting group, etc.; R<2> is a (protected) OH, etc.; R<3> to R<3> are each H, OH, etc.] such as (3S)-2-(N-tert-butyloxycarbonyl-L-tryptophyl)-1,2,3,4- tetrahydroisoquinoline-3-carboxylicacid benzyl ester. A compound of formula II (R<14> is an amino-protecting group; R<31> , R<41> , R<51> and R<61> are each H, OH, etc.) is condensed with a compound of the formula H-R<24> (R<24> is a group obtained by removing H from an amino acid) by using a condensation agent in a solvent at -30 deg.C to a room temperature to give a compound of formula III and removing the protecting group to give the component of formula I.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a pharmaceutical composition comprising a compound having a dipeptidyl peptidase IV inhibitory activity and having a tetrahydroisoquinoline skeleton as an active ingredient, a novel tetrahydroisoquinoline derivative and a method for producing the same.

[0002]

2. Description of the Related Art Dipeptidyl peptidase IV is a serine protease that specifically hydrolyzes a dipeptide of X-Pro (X may be any amino acid) from the free N-terminus of a polypeptide chain. In immune system cells, expression is induced by activation of T cells, and plays an important role in T cell activation and proliferation (European Journal of Immu).
No., 17, 1821-1826, 19
1987; Biological Chemistry Hoppe-Sailer (Biological Chemistry)
Hoppe-Seyler), Volume 371, 699-7
05, 1990). That is, when dipeptidyl peptidase IV is blocked by an antibody or an inhibitor, T
Cell activation is suppressed. In addition, there is an interest in the relationship between the present enzyme and a disease state in a collagen metabolism disorder or an immune disorder. For example, in rheumatoid patients, the dipeptidyl peptidase IV positive rate of peripheral blood T cells is increased, and high dipeptidyl peptidase IV activity is detected in urine of nephritis patients.

[0003] Examples of known dipeptidyl peptidase IV inhibitors include the tripeptide diprotin A
(L-isoleucyl-L-prolyl-L-isoleucine), diprotin B (L-valyl-L-prolyl-L-
Leucine) and diprotin C (L-valyl-L-prolyl-L-isoleucine) (JP-A-59-25366).
No.), Ala-Pro-nitrobenzoylhydroxylamine (Journal of Enzyme Inhibin)
Tion), 2, 129-142, 1988),
Ala-boroPro and Pro-boroPro
(However, boroPro indicates a compound in which the carboxyl group of proline is substituted with a B (OH) ₂ group) (Proceedings of the National Academy of Sciences of the United States. Proceedings of America
heNational Academy of Sci
encounters of the United States
of America), 88 volumes, 1556-155
9, pp. 1991) and Lys- (Z (NO ₂₎₎ - thiazolidine (however, Z (NO ₂₎ represents a 4-nitrobenzyloxycarbonyl group) (Biological Chemistry Hoppe - Sailor (Biological C
hemmistry Hoppe-Seyler),
372, pages 305-311 (1991)). However, a dipeptidyl peptidase IV inhibitor having a tetrahydroisoquinoline skeleton is not known.

[0004]

DISCLOSURE OF THE INVENTION The present invention provides a compound having an excellent dipeptidyl peptidase IV inhibitory activity and a pharmaceutical composition comprising the compound as an active ingredient.

[0005]

DISCLOSURE OF THE INVENTION The present inventors have been studying cultures of microorganisms mainly isolated from soil, and found that a compound exhibiting dipeptidyl peptidase IV inhibitory activity in a culture of Aspergillus spp. Found to be produced. These compounds were isolated and purified from the culture solution, and their physicochemical properties were examined to determine their chemical structures. As a result, they were found to be novel compounds.

[0006] On the other hand, the inventors of the present invention can specifically suppress the activation of T cells by inhibiting dipeptidyl peptidase IV, and inhibit T cells such as rheumatoid arthritis and allergy.
We thought that it would be possible to prevent and treat immune disorders and deficiencies associated with cell activation, and as a result of intensive studies, we found that a series of compounds having a tetrahydroisoquinoline skeleton, including the aforementioned microbial-derived compounds, could be used. The present invention has been completed based on the new finding that it has a peptidyl peptidase IV inhibitory effect and is effective for prevention and treatment of autoimmune diseases (arthritis, rheumatoid arthritis, etc.).

That is, the present invention is a pharmaceutical composition comprising a compound having a dipeptidyl peptidase IV inhibitory action and having a tetrahydroisoquinoline skeleton or a pharmaceutically acceptable salt thereof as an active ingredient.

Further, the present invention provides a compound represented by the general formula [I]:

[0009]

Embedded image

(Wherein R ¹ is (1) a group having a structure obtained by removing a hydroxy group of a carboxy group from an amino acid whose amino group may be protected, or (2) a protecting group of an amino group, ² is (1) a hydroxyl group which may be protected, (2) a group having a structure in which one hydrogen atom of an amino group has been removed from an amino acid whose carboxy atom group may be protected, or (3) a primary or A group having a structure in which one hydrogen atom on a nitrogen atom has been removed from a secondary amine or ammonia, and R ³ , R ⁴ , R ⁵ and R ⁶ may be the same or different and represent a hydrogen atom, a hydroxyl group or a lower alkoxy group) The novel tetrahydroisoquinoline derivatives or the pharmaceutically acceptable salts thereof, and a process for producing these compounds.

Further, the present invention provides a compound represented by the following general formula [I
I]:

[0012]

Embedded image

(Wherein, R ⁷ and R ⁸ each represent a hydroxyl group and the other represents a hydrogen atom or a hydroxyl group) or a pharmaceutically acceptable salt thereof. A compound in which R ⁷ and R ⁸ in the above general formula [II] are hydroxyl groups is hereinafter referred to as TMC-2A, and a compound in which R ⁷ is a hydroxyl group and R ⁸ is a hydrogen atom is hereinafter referred to as TMC-2B
And a compound in which R ⁷ is a hydrogen atom and R ⁸ is a hydroxyl group is hereinafter referred to as TMC-2C.

The present invention also relates to these TMC-2A, TM
This is a method for producing C-2B and TMC-2C using microorganisms.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION The pharmaceutical composition of the present invention comprises a compound having a dipeptidyl peptidase IV inhibitory activity and having a tetrahydroisoquinoline skeleton or a pharmacologically acceptable salt thereof as an active ingredient. Is raised. Specifically, dipeptidyl peptidase IV
Has inhibitory activity and the formula:

[0016]

Embedded image

Pharmaceutical compositions comprising as an active ingredient a compound having a partial structure represented by the formula or a pharmaceutically acceptable salt thereof. Furthermore, a pharmaceutical composition comprising, as an active ingredient, a tetrahydroisoquinoline derivative represented by the general formula [I] or a pharmaceutically acceptable salt thereof can be mentioned.

The pharmaceutical composition of the present invention is useful as a dipeptidyl peptidase IV inhibitor, a prophylactic / therapeutic agent for autoimmune diseases, especially a prophylactic / therapeutic agent for arthritis, and a prophylactic / therapeutic agent for rheumatoid arthritis.

The tetrahydroisoquinoline derivative of the present invention includes a compound represented by the general formula [I].

The tetrahydroisoquinoline derivative [I] of the present invention or a pharmaceutically acceptable salt thereof has excellent dipeptidyl peptidase IV inhibitory activity and is therefore useful as a dipeptidyl peptidase IV inhibitor. Furthermore, the tetrahydroisoquinoline derivative [I] or a pharmacologically acceptable salt thereof is excellent in preventing and preventing autoimmune diseases.
It has a therapeutic effect and prevents and treats autoimmune diseases, in particular, prevents and treats arthritis, and prevents and treats rheumatoid arthritis.
Useful as a therapeutic.

In the compound represented by the general formula [I],
Preferred compounds are those in which R ¹ is (1) an aryloxycarbonyl group, an aryl group-substituted lower alkoxycarbonyl group or a lower alkoxycarbonyl group, in which the amino group may be protected from the amino group, in which the hydroxy group of the carboxy group is removed. A group having a substituted structure, (2) an aryloxycarbonyl group, an aryl-substituted lower alkoxycarbonyl group or a lower alkoxycarbonyl group, a hydroxyl group in which R ² may be substituted with (1) an aryl-substituted lower alkyl group, (2 A) a group having a structure in which one hydrogen atom of an amino group has been removed from an amino acid whose carboxy group may be protected with a lower alkyl or aryl group-substituted lower alkyl, or (3) a lower alkyl or aryl-substituted lower alkyl. With one or two groups An optionally substituted amino group, R
³ , R ⁴ , R ⁵ and R ⁶ are the same or different and include a hydrogen atom, a hydroxyl group or a lower alkoxy group.

Of these, more preferred compounds include
R ¹ is (1) a benzyloxycarbonyl group or te
rt- butoxycarbonyl amino atomic group may tryptophyl group which may be protected, lysyl group or phenylalanyl group or (2) a benzyloxycarbonyl group or a tert- butoxycarbonyl group, R ² is (1) with a benzyl group A hydroxyl group which may be protected,
(2) alanine, valine, leucine, isoleucine, proline, phenylalanine, tryptophan, methionine, glycine, serine, O-benzyl-serine, threonine, cysteine, which may be protected with a carboxy atom group by a methyl group or a benzyl group; glutamine,
Asparagine, tyrosine, lysine, arginine, histidine, aspartic acid, glutamic acid and formula:

[0023]

Embedded image

Wherein R ⁷ and R ⁸ have the same meaning as described above, or a group having a structure in which one hydrogen atom of the α-amino group of the α-amino acid has been removed, or (3) t
an amino group which may be substituted with one or two groups selected from an tert-butyl group or a benzyl group, R ³ is a hydrogen atom or a lower alkoxy group, R ⁴ is a hydrogen atom or a hydroxyl group,
R ⁵ is a hydrogen atom or a lower alkoxy group, R ⁶ is a hydrogen atom,
A compound which is a hydroxyl group or a lower alkoxy group is exemplified.

Further, the compounds which are preferable in terms of pharmacological effect include:
R ¹ is a tryptophyll group, R ² is (1) a hydroxyl group, (2) alanine, valine, leucine, isoleucine, proline,
Phenylalanine, tryptophan, methionine, glycine, serine, threonine, cysteine, glutamine,
Α selected from asparagine, tyrosine, lysine, arginine, histidine, aspartic acid and glutamic acid
A group having a structure in which one hydrogen atom of an α-amino atomic group has been removed from an amino acid, or (3) an amino group, R ³ , R ⁴ ,
Compounds in which R ⁵ and R ⁶ are hydrogen atoms are mentioned.

Among these compounds, R ¹ is a tryptophyl group, R ² is (1) a hydroxyl group, (2) glutamine, serine, aspartic acid, glutamic acid, alanine, cysteine, arginine, and the like. A group having a structure in which one hydrogen atom of an α-amino atomic group has been removed from an α-amino acid selected from methionine and asparagine, or (3) an amino group, R ³ , R ⁴ , R ⁵ and R ⁶ are hydrogen atoms Compounds.

Further, as a particularly preferable compound in terms of pharmacological effect, 2-L-tryptofil-1,2,3,4-tetrahydroisoquinolyl-3-carboxylic acid can be mentioned.

Further, as another compound which is preferable in terms of efficacy, TMC-2A, TMC-2B or TMC-2
C, that is, the general formula [II]:

[0029]

Embedded image

(Wherein R ⁷ and R ⁸ have the same meanings as described above).

In the present specification, amino acids include L-form and D-form.
Body and any mixture thereof, for example, alanine, valine, leucine, isoleucine, proline, phenylalanine, tryptophan, methionine, glycine, serine, threonine, cysteine, glutamine, asparagine, tyrosine, lysine, arginine, histidine, aspartic acid And protein constituent α-amino acids such as glutamic acid, norleucine, α-aminobutyric acid, γ-
Aminobutyric acid, β-aminoisobutyric acid, β-alanine, homoserine, α-methyl-serine, O-benzyl-serine,
Aliphatic monoaminocarboxylic acids such as O-carbamyl-serine and δ-hydroxy-γ-oxo-norvaline;
Monoaminodicarboxylic acids such as aminoadipic acid, theanine, γ-methyleneglutamic acid and γ-methylglutamic acid, ornithine, β-lysine, diaminomonocarboxylic acids such as α, β-diaminopropionic acid and α, γ-diaminobutyric acid, diaminopimelin Diaminodicarboxylic acids such as acids; sulfonic acid-containing amino acids such as cysteic acid; aromatic amino acids such as thyronine, kynurenine and 3,4-dioxyphenyl-alanine; aziridine-2,3-dicarboxylic acid; 2-amino-3- (Isoxazolin-5-one-4-yl) heterocyclic amino acids such as propionic acid and anticapsin, basic amino acids such as 4-oxalidine, 4-oxolidin and 3,6-diamino-5-hydroxyhexanoic acid, cystation, lanthionine Containing sulfur and S-methyl-cysteine Amino acid, pipecolic acid, azetidine-2-carboxylic acid and 2-amino-cyclopentane-1-cyclic amino acids such as carboxylic acids, and citrulline,
Special functional group-substituted amino acids such as alanosine and azaserine, and the like;

[0032]

Embedded image

(Wherein R ⁷ and R ⁸ have the same meanings as described above).

Among them, preferred are, for example,
Alanine, valine, leucine, isoleucine, proline, phenylalanine, tryptophan, methionine,
Α-amino acids such as glycine, serine, O-benzyl-serine, threonine, cysteine, glutamine, asparagine, tyrosine, lysine, arginine, histidine, aspartic acid and glutamic acid and formulas:

[0035]

Embedded image

(Wherein, R ⁷ and R ⁸ have the same meanings as described above).

In the present specification, examples of the “group having a structure obtained by removing a hydroxy group of a carboxy atom group from an amino acid” for R ¹ include, for example, alanine, valine, leucine, isoleucine, proline, phenylalanine,
A group having a structure in which the hydroxy group of the α-carboxy group is removed from α-amino acids such as tryptophan, methionine, glycine, serine, threonine, cysteine, glutamine, asparagine, tyrosine, lysine, arginine, histidine, aspartic acid, and glutamic acid. That is, alanyl group, valyl group, leucyl group, isoleucyl group, prolyl group, phenylalanyl group, tryptoyl group, methionyl group, glycyl group, seryl group, threonyl group, cysteinyl group, glutamyl group, asparaginyl group, tyrosyl group, Examples include a lysyl group, an arginyl group, a histidyl group, an aspartyl group and a glutaminyl group. Among them, preferred examples are a tryptophyll group,
A lysyl group and a phenylalanyl group are mentioned, and a particularly preferred example is a tryptophyl group.

Examples of the “group having a structure in which one hydrogen atom of an amino group is removed from an amino acid whose carboxy group may be protected” in R ² include, for example, alanine, valine, leucine, isoleucine , Proline, phenylalanine, tryptophan, methionine, glycine, serine, threonine, cysteine, glutamine, asparagine, tyrosine, lysine, arginine, histidine, aspartic acid, glutamic acid, isoleucine methyl ester, O-benzyl-serine benzyl ester, and also a compound of the formula:

[0039]

Embedded image

(Wherein R ⁷ and R ⁸ have the same meanings as described above).
Examples include groups having a structure in which one hydrogen atom of an amino group has been removed. Of these, preferred examples include glutamine, serine, aspartic acid, glutamic acid, alanine, cysteine, arginine, a group having a structure in which one hydrogen atom of the α-amino atom group of methionine and asparagine has been removed, and more preferred examples include Examples include groups having a structure in which one hydrogen atom of the α-amino group of glutamine and serine has been removed. Another preferred example is the formula:

[0041]

Embedded image

(Wherein R ⁷ and R ⁸ have the same meanings as described above).

The “group having a structure in which one hydrogen atom on a nitrogen atom has been removed from a primary or secondary amine or ammonia” in R ² includes, for example, one group selected from lower alkyl or aryl-substituted lower alkyl. Or an amino group which may be substituted by two, and preferably an amino group which may be substituted by one or two groups selected from a tert-butyl group and a benzyl group. More specific examples thereof include a tert-butylamino group, a benzylamino group, an amino group, and the like, and more preferable examples include an amino group.

The target compound [I] of the present invention includes any of those having a tetrahydroisoquinoline skeleton in which the 3-position at the 3-position is R-configuration, S-configuration, and mixtures thereof.
Of these, compounds having the S configuration are preferred. When compound [I] further has an asymmetric carbon atom, any stereoisomer based on the asymmetric carbon atom and a mixture thereof are also included in the present invention.

Compound [I], which is an active ingredient of the present invention, comprises
Conventional methods for peptide synthesis, for example, "Peptide Synthesis" (Synthetic Chemistry Series, published by Maruzen Co., Ltd., 1975) and "Peptide Synthesis Basics and Experiments" (Maruzen Co., Ltd., 1
985) or a method analogous thereto can be produced by a liquid phase method or a solid phase method.

Among the compounds [I], a compound represented by the general formula [Ia]:

[0047]

Embedded image

(Wherein R ²² is (1) a group having a structure in which one hydrogen atom of an amino group has been removed from an amino acid whose carboxy group may be protected, or (2) a primary or secondary amine Or a group having a structure in which one hydrogen atom on a nitrogen atom has been removed from ammonia, R ¹ , R ³ ,
R ⁴ , R ⁵ and R ⁶ have the same meanings as described above), are represented by the general formula [III]:

[0049]

Embedded image

[0050] (wherein, R ¹¹ is (1) group or (2) an amino-protecting group having at least amino atomic group obtained by removing a hydroxy atomic group carboxy atomic group of amino acids are protected structure, R ³ , R ⁴ , R ⁵ and R ⁶ have the same meaning as described above) or a reactive derivative at the carboxy group thereof and a general formula [IV]:

[0051]

Embedded image

(Wherein, R ²¹ represents (1) a group having a structure in which at least one hydrogen atom of an amino group has been removed from an amino acid in which at least a carboxy group is protected, or (2) 1
With a compound having a structure in which one hydrogen atom on a nitrogen atom has been removed from a secondary or secondary amine or ammonia).

[0053]

Embedded image

Wherein R ¹¹ , R ²¹ , R ³ , R ⁴ , R ⁵ and R ⁶ have the same meaning as described above, and if necessary, the protecting group is removed. can do.

Further, among the compounds [I], the compound represented by the general formula [I-
b]:

[0056]

Embedded image

(Wherein, R ¹³ is a group having a structure in which an amino group which may be substituted with an amino group has a hydroxy group of a carboxy group removed, R ² , R ³ , R ⁴ ,
R ⁵ and R ⁶ have the same meanings as described above), are represented by the general formula [VI]:

[0058]

Embedded image

(Wherein, R ²³ represents (1) a protected hydroxyl group, and (2) a group having a structure in which at least one hydrogen atom of an amino group has been removed from an amino acid in which at least a carboxyl group has been protected. Or (3) a group having a structure in which one hydrogen atom on a nitrogen atom has been removed from a primary or secondary amine or ammonia, and R ³ , R ⁴ , R ⁵ and R ⁶ have the same meaning as described above) Compound represented by the general formula [VII]:

[0060]

Embedded image

(Wherein R ¹² represents a group having a structure in which at least the amino group is protected and the hydroxy group of the carboxy group is removed from the protected amino group) or a reactive derivative of the carboxy group thereof With the general formula [VIII]:

[0062]

Embedded image

(Wherein R ¹² , R ²³ , R ³ , R ⁴ , R ⁵ and R ⁶ have the same meaning as described above), and if necessary, the protective group is removed to prepare the compound. can do.

More specific examples of the production method are described in the following (A) to (A).
It is shown in (C).

(A) Formula [IX]:

[0066]

Embedded image

(Wherein R ¹⁴ is a protecting group for an amino group, R ³¹ ,
R ⁴¹ , R ⁵¹ and R ⁶¹ are the same or different and each represent a hydrogen atom, a hydroxyl group or a lower alkoxy group) or a reactive derivative of the carboxy group thereof and a general formula [X]:

[0068]

Embedded image

(Wherein R ²⁴ is (1) a group having a structure in which at least one hydrogen atom of an amino group is removed from an amino acid in which at least a carboxy group is protected, or (2) 1
A secondary or secondary amine or a group having a structure in which one hydrogen atom on a nitrogen atom has been removed from ammonia) using an appropriate condensing agent in an appropriate solvent.
By condensation at 30 ° C. to room temperature, the compound of the general formula [XI]:

[0070]

Embedded image

Wherein R ¹⁴ , R ²⁴ , R ³¹ , R ⁴¹ , R ⁵¹ and R ⁶¹ have the same meanings as described above, and if necessary, the protecting group is removed by a conventional method. Thereby, the corresponding target compound [I] can be produced.

Also, if desired, the compound [XI]
General formula [XII] from which terminal protecting group R ¹⁴ has been removed:

[0073]

Embedded image

Wherein R ²⁴ , R ³¹ , R ⁴¹ , R ⁵¹ and R ⁶¹
Has the same meaning as described above) and a compound of the general formula [XIII]:

[0075]

Embedded image

(Wherein R ¹⁵ represents a group having a structure in which at least an amino group in which an amino group is protected is obtained by removing a hydroxy group of a carboxy group) using an appropriate condensing agent In a suitable solvent,
Condensation is performed under ice cooling to room temperature to obtain a compound of the general formula
V]:

[0077]

Embedded image

(Wherein R ¹⁵ , R ²⁴ , R ³¹ , R ⁴¹ , R ⁵¹ and R ⁶¹ have the same meanings as described above), and if desired, a protecting group may be added according to a conventional method. By removing, the corresponding target compound [I] can be produced.

The compounds [IX], [X], [XI],
In [XII], [XIII] and [XIV], R
Preferred examples of ¹⁴ include an aryl group-substituted lower alkoxycarbonyl group, and preferred examples of R ¹⁵ include a hydroxy group of an α-carboxy group to an α-amino acid having an amino group protected by a lower alkoxycarbonyl group. Examples of R ²⁴ include a group having a structure in which one hydrogen atom of an α-amino group is removed from an α-amino acid in which a carboxy group is protected by lower alkyl. Or an amino group which may be substituted with one or two groups selected from lower alkyl or aryl-substituted lower alkyl. Examples of R ³¹ , R ⁴¹ , R ⁵¹ and R ⁶¹ are the same. Or, differently, a hydrogen atom or a lower alkoxy group can be mentioned.

(B) General formula [XV]:

[0081]

Embedded image

(Wherein, R ²⁵ represents a hydroxyl group having a protecting group, and R ³² , R ⁴² , R ⁵² and R ⁶² are the same or different and represent a hydrogen atom, a hydroxyl group or a lower alkoxy group). And a compound of general formula [XVI]:

[0083]

Embedded image

(Wherein, R ¹⁶ represents a group having a structure in which at least the amino group is protected and the hydroxy group of the carboxy group is removed from the amino acid whose amino group is protected), or a reactive derivative of the carboxy group. Is condensed with a suitable condensing agent in a suitable solvent under ice-cooling to room temperature to obtain a compound of the general formula [XVII]:

[0085]

Embedded image

Wherein R ¹⁶ , R ²⁵ , R ³² , R ⁴² , R ⁵² and R ⁶² have the same meanings as described above, and if necessary, the protecting group is removed by a conventional method. By doing so, the corresponding target compound [I] can be produced.

The compounds [XV], [XVI] and [X
VII], a preferable example of R ¹⁶ is a group having a structure in which a hydroxy atom group of an α-carboxy atom group is removed from an α-amino acid in which an amino atom group is substituted with a lower alkoxycarbonyl group. Preferred examples of ²⁵ include an aryl-substituted lower alkoxy group, and preferred examples of R ³² , R ⁴² , R ⁵² and R ⁶² include a hydrogen atom.

(C) Among the compounds obtained in the above (B), the compound represented by the general formula [XVIII]:

[0089]

Embedded image

(Wherein R ¹⁶ , R ³² , R ⁴² , R ⁵² and R ⁶²
Has the same meaning as described above) or a reactive derivative at the carboxy group thereof and a general formula [XI
X]:

[0091]

Embedded image

(Wherein R ²⁶ represents (1) a group having a structure in which at least one hydrogen atom of an amino group is removed from an amino acid in which at least a carboxy group is protected, or (2) 1
A secondary or secondary amine or a group having a structure obtained by removing one hydrogen atom on a nitrogen atom from ammonia) with a suitable condensing agent in a suitable solvent,
By condensation at −30 ° C. to room temperature, the compound represented by the general formula [X
X]:

[0093]

Embedded image

Wherein R ²⁶ , R ¹⁶ , R ³² , R ⁴² , R ⁵² and R ⁶² have the same meanings as described above, and if necessary, the protecting group is removed by a conventional method. By doing so, the corresponding target compound can be produced.

In the above-mentioned compounds [XVIII], [XIX] and [XX], preferred examples of R ¹⁶ include an α-amino acid having an amino group protected by a lower alkoxycarbonyl group to a hydroxy group of an α-carboxy atom group. A group having a structure in which an atomic group has been removed is mentioned. A preferred example of R ²⁶ is an α-amino acid in which the carboxy atomic group is protected by an aryl-substituted lower alkyl group.
A group having a structure in which one hydrogen atom of an amino group has been removed or an amino group, and R ³² , R ⁴² , R ⁵² and R
Examples of ⁶² include a hydrogen atom.

The protecting group for the carboxy group (carboxyl group) and the amino group (amino group) may be any group which does not participate in the condensation reaction and can be easily removed by a conventional method. What is usually used can be used. Examples of the protecting group for the carboxy atom group include a lower alkyl group and an aryl group-substituted lower alkyl group. Specific examples include a methyl group, an ethyl group, and a benzyl group. Among them, preferred are aryl-substituted lower alkyl groups, such as benzyl group. Examples of the protecting group for the amino group include a substituted and unsubstituted lower alkoxycarbonyl group, and specific examples thereof include a benzyloxycarbonyl group, a 4-methoxybenzyloxycarbonyl group, and a 9-fluorenylmethyloxycarbonyl group. A tert-butoxycarbonyl group and 2,
And a 2,2-trichloroethyloxycarbonyl group. Among them, preferred are an aryl-substituted lower alkoxycarbonyl group and an unsubstituted lower alkoxycarbonyl group, such as a benzyloxycarbonyl group and a tert-butoxycarbonyl group.

The protecting groups for the carboxy and amino groups can be easily removed by a known method, for example, a conventional method of peptide chemistry.

Examples of the reactive derivative at the carboxyl group of the amino acid include its active ester, such as succinimide ester and benzotriazole ester.

As a suitable condensing agent, 1,3-dicyclohexylcarbodiimide and the like are used. The condensation reaction can also be carried out using a combination of an ester activator and a condensing agent, for example, 1-hydroxybenzotriazole (monohydrate) and 1-ethyl-3- (3-dimethylaminopropyl) carboxydiimide hydrochloride. Salts, N-hydroxysuccinimide and 1,3-dicyclohexylcarbodiimide can be used. Of these, a combination of 1-hydroxybenzotriazole (monohydrate) and 1-ethyl-3- (3-dimethylaminopropyl) carboxydiimide hydrochloride is preferred.

Suitable solvents may be any inert solvents which do not participate in the condensation reaction, such as dimethylformamide, dimethylsulfoxide, dichloromethane, dichloroethane, chloroform, tetrahydrofuran, ethyl acetate and N-methylpyrrolidone. Preferred is dimethylformamide.

When the compound to be subjected to the condensation reaction has a reactive functional group other than the carboxy atom group and the amino atom group, the functional group is protected in advance and then subjected to the condensation reaction according to a conventional method. It is preferable to appropriately deprotect.

The target compound [I] of the present invention is subjected to condensation and deprotection of a carrier obtained by binding a desired starting amino acid to a resin and a corresponding starting amino acid derivative by using a commercially available automatic synthesizer. After removing the resin, means for separating the peptides, such as extraction, distribution, reprecipitation, crystallization,
It can also be obtained by purification by recrystallization, various types of chromatography, high-performance chromatography and the like.

As the resin, any resin can be used as long as the target substance can be finally cut out in the form of an amide. For example, N-α-9-fluorenylmethoxycarbonyl-superacid dradium can be used. Building Polyethylene glycol Handle Polystyrene (Product name: Fm
oc-NH-SAL-PEG Resin; manufactured by Watanabe Chemical), (4-2 ′, 4′-dimethoxyphenyl-N-α)
-9-Fluorenylmethoxycarbonyl-aminomethyl) -phenoxy resin (trade name: Fmoc-NH-
SAL Resin; manufactured by Watanabe Chemical) and (4-2 ',
4'-dimethoxyphenyl-N-α-9-fluorenylmethoxycarbonyl-aminomethyl) -phenoxyacetamide-norleucine-p-methyl-benzhydroxyamine resin (trade name: Fmoc-NH-SAL-
MBHA Resin (manufactured by Watanabe Chemical).

Further, among the compounds which are the active ingredients of the present invention, general formula [II]:

[0105]

Embedded image

(Wherein R ⁷ and R ⁸ have the same meanings as described above), that is, TMC-2A, TMC-2B or TMC-2C
Can also be obtained by culturing mold belonging to the genus Aspergillus and isolating it from the culture.

The method for producing TMC-2A, TMC-2B and TMC-2C by a mold belonging to the genus Aspergillus will be described in more detail below.

TMC-2A, TMC-2B and TMC
An example of a 2C-producing bacterium is A374 strain isolated from soil in Kochi City, Kochi Prefecture. The bacteriological properties of this strain are as follows.

The A374 strain was incubated at 25 ° C.
Table 1 shows the growth state after culturing for one day. The color tone was determined according to the JIS standard color table (Z8721).

[0110]

[Table 1]

The growth temperature range of strain A374 is 15 to 45 ° C.
And the optimal temperature range is 20 to 40 ° C. The growth pH range is pH 2 to 13, and the optimal pH range is p.
H3 to H11.

The mycelium has a smooth surface and partition walls. From part of the hypha, foot cells (foot Cell;
Conidia stalks (6.7-8.0 μm × 23-1100 μm) with 6.7-8.7 μm × 37-57 μm) are formed, and the subcapsular, partially club-shaped apical sac (17) is formed at the tip. ~ 2
4 μm × 19-27 μm). A pin-shaped primary incision (primary) from the upper semicircular portion of the apical sac
steigmata; 2.3 to 3.3 μm × 13 μ
m), from which spherical conidia (4.0-6.7μ)
m) are formed in a chain or in a chain in a chain. The conidial head is mainly cylindrical (50-100 μm × 160-200).
μm), but globe-like or spherical (33-60 μm ×
Conidia heads having a shape of 20 to 47 µm) are also observed.

From the above characteristics, it was found that the A374 strain belongs to the genus Aspergillus. This Aspergillus sp. A374 strain was submitted to the Research Institute of Biotechnology, Industrial Technology Institute of the Ministry of International Trade and Industry on May 18, 1995 under the accession number FERM P-14934.
And deposited on September 19, 1997 under the accession number FERM BP-6113.

According to the method of the present invention, TMC-2A, TM
To produce C-2B and TMC-2C, TMC-2A, TMC-2B and TMC belonging to the genus Aspergillus are used.
MC-2C-producing bacteria are inoculated into a nutrient-containing medium and grown aerobically. Thereby, TMC-2A, TMC-2
A culture containing B and TMC-2C is obtained.

As a nutrient, a carbon source and a nitrogen source that can be used as nutrients for microorganisms can be used.
For example, peptone, meat extract, corn steep
Liquor, cottonseed flour, peanut flour, soy flour, yeast extract, NZ
-Nitrogen sources such as amines, casein hydrolysates, ammonium nitrate, ammonium sulfate and carbon sources such as carbohydrates or fats such as starch, glycerin, sucrose, glucose, galactose, mannose, molasses can be used. Also, salt, calcium carbonate, phosphate,
Inorganic salts such as magnesium sulfate can be added. These are used by producing bacteria, and TMC-2A, TMC-2B
And TMC-2C may be used.

The TMC-2A, TMC-2B and T
Liquid culture is preferable for culture of MC-2C-producing bacteria. The cultivation temperature can be used in a range in which the producing bacteria grow and produce a desired substance, and is usually 20 to 35 ° C. The cultivation can be carried out by appropriately selecting from the above conditions according to the properties of the producing bacteria to be used. The desired TMC-2A, TMC-2B and TMC-2C are produced in the culture. Isolation and purification of these products can be performed by methods known per se, for example, ion exchange chromatography, partition chromatography,
Reverse phase chromatography and the like can be appropriately combined.

Dipeptidyl Peptidase IV Inhibitors of the Invention TMC-2A, TMC-2B and TMC-2C
Is shown in Table 2 below.

[0118]

[Table 2]

Further, TMC-2A, TMC-2B and TMC-2B
The UV and IR spectra of MC-2C are shown in the attached FIGS. 1-3 and 4-6, respectively. The UV spectrum was analyzed on a 50 μg / ml methanol solution of each sample. IR spectra were analyzed on potassium bromide tablets containing 1% (w / w) of each sample.

Further, the 400 MHz proton nuclear magnetic resonance spectra of TMC-2A, TMC-2B and TMC-2C are shown in FIGS. Note that TMC
-2A and TMC-2B were measured in heavy water using TSP (sodium trimethylsilylpropylsulfonate) as an internal standard. TMC-2C was measured in heavy methanol using TMS (tetramethylsilane) as an internal standard. The chemical shifts δ (ppm) are described below.

TMC-2A: 7.55 (1H, d),
7.46 (1H, d), 7.34 (1H, s), 7.1
3 (1H, t), 7.07 (1H, t), 6.02 (1
H, s), 4.82 (1H, d), 4.50 (1H, d
d), 4.11 (1H, dd), 3.77 (1H,
d), 3.73 (4H, m), 3.49-3.28 (5
H, m), 3.19 (1H, dd), 2.35 (1H,
dd), 1.68 (1H, ddd), 1.44 (1H,
ddd), 1.14 (1H, dd), 0.71 (1H,
m) TMC-2B: 7.58 (1H, dd), 7.52 (1
H, d), 7.38 (1H, s), 7.22 (1H, d
d), 7.16 (1H, dd), 6.09 (1H,
s), 4.80 (1H, d), 4.50 (1H, d)
d), 4.04 (1H, dd), 3.83 (1H,
d), 3.77 (1H, m), 3.73 (3H, s),
3.50 (1H, dd), 3.43 (1H, dd),
3.08 (2H, d), 2.39 (1H, dd), 1.
64 (1H, ddd), 1.20 (2H, m), 0.6
3 (4H, m) TMC-2C: 7.53 (1H, d), 7.40 (1
H, d), 7.24 (1H, s), 7.12 (1H, d
d), 7.07 (1H, dd), 6.09 (1H,
s), 5.06 (1H, d), 4.23 (1H, d
d), 4.22 (1H, dd), 3.73 (3H,
s), 3.70 (1H, d), 3.60 (1H, d)
d), 3.36 (2H, d), 3.12 (2H, d),
2.37 (1H, dd), 1.45 (2H, dd),
1.29 (1H, m), 0.58 (1H, m), 0.5
0 (3H, d) 10 of TMC-2A, TMC-2B and TMC-2C
The 0 MHz carbon nuclear magnetic resonance spectra are shown in FIGS. In addition, TMC-2A and TMC
-2B was measured in heavy water using dioxane as an internal standard. TMC-2C was measured in heavy methanol using TMS as an internal standard. Also, their chemical shift δ
(Ppm) are described below.

TMC-2A: 181.4, 174.1,
173.7, 151.1, 148.6, 139.6, 1
37.1, 132.0, 129.2, 128.0, 12
5.2, 122.6, 120.7, 114.9, 11
3.6,109.9,109.1,65.6,63.
5,62.7,59.3,55.5,54.8,41.
6,41.4,33.1,32.2,30.5 TMC-2B: 182.0,174.4,174.2,
151.5, 149.1, 139.5, 137.7, 1
32.4, 129.6, 128.5, 125.7, 12
3.1, 121.2, 115.4, 114.0, 11
0.4, 109.1, 67.6, 64.0, 59.7,
56.3, 55.2, 42.1, 37.7, 34.4,
33.4, 30.9, 20.1 TMC-2C: 179.6, 171.9, 171.4,
150.8, 148.0, 138.1, 135.8, 1
30.0, 128.1, 125.6, 123.2, 12
0.6, 119.3, 112.7, 111.4, 10
8.2, 107.7, 68.6, 60.9, 58.2
53.6, 53.5, 39.9, 36.9, 33.1,
31.7, 29.5, 15.5 The protons bonded to the nitrogen atom and the oxygen atom were prepared by preparing an acetylated form of TMC-2A, and measuring the proton and carbon nuclear magnetic resonance spectra in deuterated chloroform. Confirmed by measurement.

On the basis of the above results, TMC-2A,
TMC-2B and TMC-2C have the general formula [I
I]. No compound of this chemical structure has been reported so far, and TMC-2A,
TMC-2B and TMC-2C are new substances.

[0124] Whether a compound has a dipeptidyl peptidase IV inhibitory activity is determined by, for example, determining whether the compound has an L-G
ly-L-Pro-p-nitroanilide is L-Gly-
The determination can be made based on whether or not the reaction hydrolyzed by L-Pro and p-nitroaniline is inhibited.

The compounds of the present invention can be used for pharmaceutical use in free form or in the form of a pharmaceutically acceptable salt. Such pharmacologically acceptable salts may be any conventional non-toxic salts, for example, inorganic acid salts such as hydrochloride, hydrobromide, sulfate or phosphate, and formate. , Acetate, trifluoroacetate, oxalate, maleate, fumarate, tartrate, metasulfonate, benzenesulfonate, toluenesulfonate, and other organic acid salts, sodium salts, potassium salts, etc. And alkaline earth metal salts such as calcium salts and the like, and salts with amino acids such as arginine salts, aspartates and glutamates.

The compounds of the present invention and pharmaceutically acceptable salts thereof are to be construed as including any of inner salts, addition salts, solvates, hydrates and the like. .

When a pharmaceutical composition comprising the compound of the present invention and a pharmaceutically acceptable salt thereof as an active ingredient is used as a drug, an aerosol, tablet, pill, powder, granule,
Lozenges, solutions, suspensions, emulsions, capsules, microcapsules, suppositories, injections, plasters, ointments, syrups, cataplasms, liniments, lotions, and other conventional pharmaceutical preparations Orally or parenterally (intravenously, intramuscularly, intradermally, subcutaneously, intraperitoneally or rectally).

[0128] Additives for these drugs may be used as long as they do not impair the therapeutic effect of each drug and are harmless in the dose of the drug. Any conventional additives can be used. For example, stabilizers, buffers, flavoring agents, suspending agents, emulsifiers, fragrances, preservatives, solubilizers, excipients, coloring agents, binders, disintegrants, sweeteners, thickeners, wetting Agents, solvents and the like can be used.

The amount of the active ingredient in the pharmaceutical preparation may be an amount sufficient to produce the desired therapeutic effect, and is, for example, 0.01 mg / kg to 100 mg / kg for oral or parenteral administration, preferably 1 mg / kg. / Kg to 30 mg / kg.

In the present specification, examples of the protecting group for the amino group (the protecting group for the amino group) in the “amino acid whose amino group may be protected” include, for example, an acyl group, an akanoyl group, an aroyl group, an aralkyl group. Examples include a carbonyl group, an aryloxycarbonyl group, an aryl group-substituted lower alkoxycarbonyl group or a lower alkoxycarbonyl group, and particularly an aryloxycarbonyl group, an aryl group-substituted lower alkoxycarbonyl group or a lower alkoxycarbonyl group. The amino acid having a protected carboxy group in the “amino acid whose carboxy group may be protected” is a compound in which the carboxy group of the amino acid is esterified (amino acid ester) or the carboxy group of the amino acid is amidated. Examples of such compounds include amino acids in which the carboxy group is protected with a lower alkyl, amino acids in which the carboxy group is protected with an aryl-substituted lower alkyl, and carboxy group protected with a di-lower alkylamine. Amino acids and the like.

When the “amino acid” has a reactive residue other than the amino group and the carboxy group (for example, a hydroxyl group in serine), the peptide is synthesized according to the type of the reactive residue. May be protected with a protecting group generally used in the field of (for example, a benzyl group in the case of a hydroxyl group).

The "aryloxycarbonyl group" includes a phenoxycarbonyl group and a naphthyloxycarbonyl group. The "aryl group-substituted lower alkoxycarbonyl group" includes a benzyloxycarbonyl group, a phenethyloxycarbonyl group and a naphthylmethyloxy group. And preferably a benzyloxycarbonyl group. Further, as the "lower alkoxycarbonyl group", methoxycarbonyl group, ethoxycarbonyl group, n-propoxycarbonyl group, isopropoxycarbonyl group, n-butoxycarbonyl group, isobutoxycarbonyl group, tert-butoxycarbonyl group, s
ec-butoxycarbonyl group, n-pentylcarbonyl group, isopentylcarbonyl group, sec-pentylcarbonyl group, tert-pentylcarbonyl group, n-hexylcarbonyl group, isohexylcarbonyl group, sec
-Hexylcarbonyl group and tert-hexylcarbonyl group, and the like, and preferably a tert-butoxycarbonyl group. Examples of the “aryl-substituted lower alkyl” include a benzyl group, a phenethyl group and a naphthylmethyl group, preferably benzyl.

In the present specification, aryl means phenyl, naphthyl and the like. In addition, lower alkyl and lower alkoxy represent a branched or straight chain having 1 to 6 carbon atoms, preferably a branched or straight chain having 1 to 4 carbon atoms.

[0134]

EXAMPLES The present invention will be described more specifically with reference to the following experimental examples and examples, but the present invention is not limited to these examples.

Experimental Example 1 The reaction was performed according to Nagatsu et al. (Analytical Biochemistry).
y), Vol. 74, pp. 466-476, 1976) using a 96-well flat bottom plate at 37 ° C.
Cider et al. (Preparative Biochemistry (P
reparative Biochemistry),
21 (2-3), 141-150, 1991), a dipeptidyl peptidase IV enzyme solution (25 mU / ml) derived from Lewis rat kidney 5
μl, 30 μl of water and 5 μl of a 2 mM dimethyl sulfoxide solution of the test compound, and pre-incubation was performed for 10 minutes, and then 710 mM Gly-Na
OH (pH 8.7) buffer solution 10 μl, 3 mM LG
ly-L-Pro-p-nitroanilide (Gly-Pr
o-pNA (manufactured by Sigma) aqueous solution (50 μl) was added, and the amount of p-nitroaniline generated was measured using a plate reader (T
By measuring the increase in absorbance (ΔOD) at a wavelength of 405 nm using HERMOmax (manufactured by Molecular Devices), the inhibition rate (%) of dipeptidyl peptidase IV enzyme activity is determined by the following formula 1. However, 1 U of dipeptidyl peptidase IV enzyme activity is 1 μm per minute.
It is the amount of enzyme that produces mol of p-nitroaniline.
The results are as shown in Table 3.

[0136]

(Equation 1)

[0137]

[Table 3]

Experimental Example 2 As in Experimental Example 1, 3 mM Gly-Pro-pNA5
After adding 0 μl and 10 μl of the sample and keeping the mixture at 37 ° C. for 15 minutes, 10 ml of a 710 mM glycine buffer (pH 8.7) was added.
μl, 25 μl of distilled water and dipeptidyl peptidase IV enzyme from Lewis rat kidney (50 mU / m
l) 5 μl of the solution was added and mixed, and reacted at 37 ° C., and the inhibition rate (%) of dipeptidyl peptidase IV enzyme activity was determined by the above formula 1. As a result, TM of 4.6 μg / ml
C-2A, 9.5 μg / ml TMC-2B or 11
μg / ml of TMC-2C inhibited dipeptidyl peptidase IV activity by 50%.

TMC-2A, TMC-2B and TMC
It was confirmed that -2C also inhibited dipeptidyl peptidase IV prepared from rat spleen, human peripheral blood mononuclear cells and human colon cancer cell line Caco2, in addition to dipeptidyl peptidase IV purified from rat kidney.

Furthermore, the effect of TMC-2A, TMC-2B and TMC-2C on other peptidases was examined. That is, at a concentration of 100 μg / ml, TM
The effects of C-2A, TMC-2B and TMC-2C on prolyl endopeptidase, subtilisin, trypsin, cathepsin C, leucine aminopeptidase and proline aminopeptidase were examined. As a result, TMC-2A and TMC-2B had no effect on all the tested peptidases. On the other hand, TMC-2C
Showed weak inhibition of prolyl endopeptidase and proline aminopeptidase, but did not inhibit other peptidases. Therefore, TMC-2A, TM
C-2B and TMC-2C were found to be highly specific inhibitors of dipeptidyl peptidase IV.

Experimental Example 3 By administering 35 mg / kg of alkyldiamine to the ridge of a F344 / Jcl rat (five weeks old, female),
Arthritis was induced. TMC-2A was dissolved in physiological saline, and for 3 weeks from the date of alkyldiamine administration to the end of the test,
It was administered once daily subcutaneously on the back. The dose is 30, 10,
3 and 1 mg / kg.

FIG. 13 shows how much the footpad swelled at the end of the test in each test group (percentage of the footpad swelling based on the volume of the footpad before alkyldiamine administration in each individual). In addition, the volume of the footpad was measured using a foot volume measuring device Pletysmometer (manufactured by Unicom). As is clear from FIG. 13, TMC-2A suppressed the onset and progression of alkyldiamine-induced arthritis in a dose-dependent manner. In particular, 30 mg /
In the administration groups of 10 kg / kg and 10 mg / kg, the suppression was significantly suppressed as compared with the control group (the physiological saline administration group).

Experimental Example 4 Heat-killed Mycobacterium tuberculosis (M. tubercu) suspended in liquid paraffin at the ridge of a Lewis rat (6 weeks old, female)
0.6 mg of losis H37Ra strain (manufactured by Difco)
/ Kg was administered to induce arthritis. The test compound was dissolved in physiological saline and administered subcutaneously on the back once a day for 18 days from the administration of the heat-killed Mycobacterium tuberculosis to the end of the test. The dose is 10 mg / mg for TMC-2A.
kg, 30 and 10 mg for the compound of Example 17
/ Kg.

FIG. 14 shows how much the footpad swelled in each test group at the end of the test (percentage of the footpad swelling based on the volume of the footpad before administration of the heat-killed M. tuberculosis in each individual). Show. In addition, the volume of the footpad was measured using a foot volume measuring device Pletysmometer (manufactured by Unicom). As is clear from FIG. 14, TMC-2A and the compound of Example 13 suppressed the onset and progression of adjuvant-induced arthritis in a dose-dependent manner. In particular, 10 mg / kg of TMC-2A, Example 13
In the group administered with 30 mg / kg of the compound (1), the compound was significantly suppressed as compared with the control group (group administered with physiological saline).

Example 1 7.3 g of (3S) -1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid / benzyl ester, Nt
4.56 g of tert-butyloxycarbonyl-L-tryptophan, 2.76 g of 1-hydroxybenzotriazole (monohydrate) and 3.45 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride were
After dissolving in 50 ml of dimethylformamide under ice-cooling and stirring for 2 hours under ice-cooling and further for 16 hours at room temperature, the reaction solution was concentrated under reduced pressure. Extract the residue with ethyl acetate, wash, dry,
Concentrated. The residue was purified by silica gel column chromatography (ethyl acetate: n-hexane = 1: 2) to obtain amorphous (3S) -2- (N-tert-butyloxycarbonyl-L-tryptofil) -1,2,2,3.
3,4-tetrahydroisoquinoline-3-carboxylic acid
8.07 g of the benzyl ester was obtained. IR (KBr, cm ^-1 ): 3320, 2975, 173
0, 1700, 1645, 1450, 1430, 117
0,745 MS (SIMS): 554 (M + 1) Examples 2-6 Performed 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid benzyl ester and N-tert-butoxycarbonyl-α-amino acid derivative The same treatment as in Example 1 was carried out to obtain the compounds shown in Table 4 below.

[0146]

[Table 4]

Example 7 7.0 g of the compound obtained in Example 1 was added to 50 ml of methanol.
After addition of a catalytic amount of palladium-carbon, the solution was hydrogenated at room temperature under balloon pressure for 3 hours. The catalyst was filtered off and the filtrate was concentrated. The residue was crystallized from an ethyl acetate-n-hexane solvent to give colorless crystals of (3S) -2.
5.25 g of-(N-tert-butyloxycarbonyl-L-tryptofil) -1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid was obtained. m. p. 143 ° C (decomposition) IR (KBr, cm ^-1 ): 3340, 2980, 172
0, 1700, 1630, 1440, 1170, 740 MS (SIMS): 464 (M + 1) Examples 8 to 11 The compounds obtained in Examples 2 to 5 were treated in the same manner as in Example 7 and described in Table 5 below. The compound was obtained.

[0148]

[Table 5]

Example 12 2.97 g of the compound obtained in Example 6 was treated with methanol 20
The mixture was dissolved in a 1 ml aqueous solution, and 5 ml of a 1 M aqueous sodium hydroxide solution was added thereto, followed by stirring at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the residue was washed with ether. The mixture was acidified with an aqueous potassium hydrogen sulfate solution, and extracted with ethyl acetate. The extract is washed, dried and concentrated to obtain an amorphous (3S) -2-
｛N (α) -tert-butyloxycarbonyl-N
(Ω) -benzyloxycarbonyl-L-lysyl}-
2.62 g of 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid was obtained. IR (KBr, cm ^-1 ): 3340, 2975, 170
5,1625,1520,1440,1240,116
5 MS (SIMS): 540 (M + 1) Example 13 4.0 g of the compound obtained in Example 7 was dissolved in 50 ml of a 4M hydrochloric acid-dioxane solution, and the mixture was dissolved in a nitrogen atmosphere at room temperature for 30 minutes.
Stirred for minutes. The reaction solution was concentrated under reduced pressure, the residue was dissolved in water, and then freeze-dried to give a pale red powder (3S) -2-
3.3 g of (L-tryptofil) -1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid hydrochloride was obtained. m. p. : 158 ° C (decomposition) IR (KBr, cm ^-1 ): 3400, 2910, 172
0, 1645, 1490, 1460, 1205, 112
0,745 MS (SIMS): 364 (M + 1) Examples 14 to 18 The compounds obtained in Examples 8 to 12 were treated in the same manner as in Example 13 to obtain the compounds shown in Table 6 below.

[0150]

[Table 6]

[0151]

[Table 7]

Example 19 463 mg of the compound obtained in Example 7 and O-benzyl-
L-serine benzyl ester 410 mg, 1-hydroxybenzotriazole (monohydrate) 184 mg, 1-hydroxybenzotriazole (monohydrate)
Ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (230 mg) was dissolved in dimethylformamide (20 ml) under ice cooling, and the mixture was stirred for 2 hours under ice cooling and further at room temperature for 16 hours, and then concentrated under reduced pressure. The residue was extracted with ethyl acetate, washed, dried and concentrated. The residue was subjected to silica gel column chromatography (ethyl acetate: n-hexane =
Purified by 1: 1), amorphous N-｛(3S)
-2- (N-tert-butyloxycarbonyl-L-
670 mg of (tryptofil) -1,2,3,4-tetrahydroisoquinoline-3-carbonyl} -O-benzyl-L-serine benzyl ester were obtained. IR (KBr, cm ^-1 ): 3305, 2975, 173
5,1660,1640,1480,1450,117
0,745 MS (SIMS): 731 (M + 1) Example 20 The compound obtained in Example 7 and L-leucine benzyl ester were treated in the same manner as in Example 19 to give N-｛(3S).
-2- (N-tert-butyloxycarbonyl-L-
(Triptofil) -1,2,3,4-tetrahydroisoquinoline-3-carbonyl} -L-leucine benzyl ester was obtained. m. p. MS (SIMS): 667 (M + 1) Example 21 The compound obtained in Example 19 was treated in the same manner as in Example 7, and the product was subjected to silica gel chromatography (chloroform: methanol = 20). 1) and separated in (1) amorphous N-｛(3S) -2- (N-tert-butyloxycarbonyl-L-tryptofil) -1,
2,3,4-tetrahydroisoquinoline-3-carbonyl} -L-serine and (2) amorphous N-
{(3S) -2- (N-tert-butyloxycarbonyl-L-tryptofil) -1,2,3,4-tetrahydroisoquinoline-3-carbonyl} -O-benzyl-L-serine was obtained. (1) IR (KBr, cm ^-1 ): 3315, 2960, 168
0, 1640, 1500, 1450, 1165, 745 MS (SIMS): 641 (M + 1) (2) IR (KBr, cm ^-1 ): 3335, 2960, 168
0,1635,1510,1450,1430,116
5,745 MS (SIMS): 551 (M + 1) Example 22 1.16 g of the compound obtained in Example 20 was dissolved in 20 ml of a 4M hydrochloric acid-dioxane solution, and the mixture was stirred under a nitrogen atmosphere at room temperature for 30 minutes. The reaction solution was concentrated under reduced pressure, and the residue was crystallized from ethanol to give N- 無色 (3S) -2 as a colorless powder.
990 mg of -L-tryptofil-1,2,3,4-tetrahydroisoquinoline-3-carbonyl} -L-leucine benzyl ester were obtained. m. p. : 132-134 ° C MS (SIMS): 567 (M + 1) Example 23 N-{(3S) -2- (N-tert-butyloxycarbonyl-L-tryptofil) -1,2,3,4-tetrahydroisoquinoline -3-Carbonyl} -L-serine was treated in the same manner as in Example 13 to give amorphous N-
｛(3S) -2-L-tryptofil-1,2,3,4
-Tetrahydroisoquinoline-3-carbonyl} -L-
Serine hydrochloride was obtained. IR (KBr, cm ^-1 ): 3400, 2940, 172
5,1650,1450,745 MS (SIMS): 451 (M + 1) Example 24 750 mg of the compound obtained in Example 22 was added to methanol 5
After dissolving in 0 ml and adding a catalytic amount of palladium-carbon, the solution was hydrogenated at room temperature under balloon pressure for 3 hours.
The catalyst was filtered off, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was crystallized from ethanol to give colorless crystals of N-｛(3S).
650 mg of -2-L-tryptofil-1,2,3,4-tetrahydroisoquinoline-3-carbonyl} -L-leucine hydrochloride was obtained. m. p. MS (SIMS): 477 (M + 1) Example 25 (3S) -2-benzyloxycarbonyl-5,8-dimethoxy-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid 4 1.5 g, L-leucine methyl ester hydrochloride 1.82 g, 1-hydroxybenzotriazole (monohydrate) 1.95 g, 1-ethyl-3- (3-
Dimethylaminopropyl) carbodiimide hydrochloride 2.4
4 g and 1.7 ml of triethylamine were dissolved in 35 ml of dimethylformamide under ice-cooling, and the mixture was stirred under ice-cooling for 2 hours and at room temperature for 16 hours, and then concentrated under reduced pressure. The residue was extracted with ethyl acetate, washed, dried and concentrated. The residue was purified by silica gel column chromatography (chloroform: ethyl acetate = 9: 1) to give a colorless oil of N-｛(3
S) -2-Benzyloxycarbonyl-5,8-dimethoxy-1,2,3,4-tetrahydroisoquinoline-3
-Carbonyl｝ -L-leucine methyl ester 5.0
g was obtained. IR (KBr, cm ^-1 ): 3400, 2955, 174
0, 1670, 1485, 1260, 1120, 108
5 MS (SIMS): 499 (M + 1) Example 26 700 mg of the compound obtained in Example 25 was added to methanol 1
0 ml, 1 M sodium hydroxide aqueous solution 2.8 m
After adding l, the mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the residue was washed with ether. The mixture was acidified with an aqueous potassium hydrogen sulfate solution, and extracted with ethyl acetate. The extract was washed, dried and concentrated to obtain amorphous N-｛(3
S) -2-Benzyloxycarbonyl-5,8-dimethoxy-1,2,3,4-tetrahydroisoquinoline-3
470 mg of -carbonyl｝ -L-leucine were obtained. IR (KBr, cm ^-1 ): 3400, 2960, 168
5,1485,1260,1085 MS (SIMS): 485 (M + 1) Example 27 4.0 g of the compound obtained in Example 25 was converted to methanol 40.
After dissolving in p.m. and adding a catalytic amount of palladium-carbon,
The solution was hydrogenated under balloon pressure at room temperature for 3 hours. The catalyst was filtered off, and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (chloroform: ethyl acetate = 2: 1), and the obtained colorless oil was crystallized from isopropyl ether to give N-colorless crystals.
2.02 g of ((3S) -5,8-dimethoxy-1,2,3,4-tetrahydroisoquinoline-3-carbonyl) -L-leucine methyl ester was obtained. Melting point: 101-115 ° C IR (KBr, cm ^-1 ): 3360, 2955, 173
5,1665,1480,1255,1075 MS (SIMS): 365 (M + 1) Example 28 1.5 g of the compound obtained in Example 27, 1.39 g of N-benzyloxycarbonyl-L-tryptophan, 1-
Hydroxybenzotriazole (monohydrate) 662m
g, 1-ethyl-3- (3-dimethylaminopropyl)
After dissolving 828 mg of carbodiimide hydrochloride in 15 ml of dimethylformamide under ice cooling, the mixture was stirred for 2 hours under ice cooling and further at room temperature for 16 hours, and then the reaction solution was concentrated under reduced pressure. The residue was extracted with ethyl acetate, washed, dried and concentrated. The crystalline residue was crystallized from ethyl acetate to give colorless crystals of N-｛(3
S) -2- (N-Benzyloxycarbonyl-L-tryptofil) -5,8-dimethoxy-1,2,3,4-
2.20 g of tetrahydroisoquinoline-3-carbonyl} -L-leucine methyl ester were obtained. m. p. : 130-149 ° C IR (KBr, cm ^-1 ): 3315, 2955, 174
0, 1660, 1530, 1485, 1260, 108
5,745 MS (SIMS): 685 (M + 1) Example 29 350 mg of the compound obtained in Example 28 was added to methanol 1
After dissolving in 5 ml and adding a catalytic amount of palladium-carbon, the solution was hydrogenated at room temperature under balloon pressure for 3 hours.
The catalyst was filtered off, and the filtrate was concentrated under reduced pressure to obtain a crystalline residue.
The crystalline residue was washed with isopropyl ether to obtain amorphous N-Ｎ (3S) -2-L-tryptofil-
170 mg of 5,8-dimethoxy-1,2,3,4-tetrahydroisoquinoline-3-carbonyl} -L-leucine methyl ester was obtained. IR (KBr, cm ^-1 ): 3295, 2955, 174
0, 1645, 1485, 1260, 1090, 745 MS (SIMS): 551 (M + 1) Example 30 1.3 g of the compound obtained in Example 28 was converted to methanol 30.
dissolve in 1 ml of water, 3.8 ml of 1 M aqueous sodium hydroxide solution
Was added, and the mixture was stirred at room temperature for 6 hours. The reaction solution was concentrated under reduced pressure, the residue was washed with ether, neutralized with an aqueous solution of potassium hydrogen sulfate, and extracted with ethyl acetate. The extract was washed, dried and concentrated to obtain a crystalline residue. The crystalline residue was washed with isopropyl ether to give N-
｛(3S) -2 (N-benzyloxycarbonyl-L-
Tryptofil II-5,8-dimethoxy-1,2,3
4-tetrahydroisoquinoline-3-carbonyl) -L
-1.11 g of leucine were obtained. m. p. : 152-166 ° C IR (KBr, cm ^-1 ): 3310, 2960, 170
0, 1675, 1520, 1485, 1260, 108
5,745 MS (SIMS): 671 (M + 1) Example 31 600 mg of the compound obtained in Example 30 was treated with methanol 1
After dissolving in 5 ml and adding a catalytic amount of palladium-carbon, the solution was hydrogenated at room temperature under balloon pressure for 3 hours.
The catalyst was filtered off, and the filtrate was concentrated under reduced pressure to obtain a crystalline residue.
The crystalline residue was washed with isopropyl ether to obtain amorphous N-Ｎ (3S) -2-L-tryptofil-
5,8-Dimethoxy-1,2,3,4-tetrahydroisoquinoline-3-carbonyl} -L-leucine 345m
g was obtained. IR (KBr, cm ^-1 ): 3400, 2955, 165
5, 1480, 1260, 1085, 745 MS (SIMS): 537 (M + 1) Examples 32 to 51 PPSSM-8, an automatic peptide synthesizer with multiple solid-phase methods simultaneously.
(Manufactured by Shimadzu Corporation), and peptide synthesis was performed according to the following procedure. A carrier (Fmoc) in which the corresponding raw material amino acid is bonded to a benzoxybenzyl alcohol type resin
-AminoAcid-Resin) 100 mg, and benzotriazol-1-yl-oxy-tris (pyrrolidino) phosphonium hexafluorophosphate, 1-hydroxybenzotriazole as a condensation system,
The system of N-methylmorpholine, 20% piperidine / N, N-dimethylformamide as a deprotection system, and N-α-9-fluorenylcarbonyl- as a condensed amino acid
1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid and N-tert-butyloxycarbonyl-L-
The synthesis was performed using tryptophan using standard protocols provided with the synthesizer. After the synthesis, the mixture was treated with 1 ml of trifluoroacetic acid: water: thioanisole: ethanedithiol (75: 10: 10: 5) for 3 hours, deprotected and cleaved from the resin. The reaction solution was filtered to remove the resin, and the filtrate was precipitated by adding anhydrous diethyl ether, or concentrated to obtain the desired crude peptide.
The obtained crude peptide was dissolved in water, purified through a short reversed-phase column, and then freeze-dried to give a colorless powder of N-｛(3S) -2-L-tryptofil-1, as shown in Table 7.
2,3,4-Tetrahydroisoquinoline-3-carbonyl) -L-amino acid was obtained.

[0153]

[Table 8]

[0154]

[Table 9]

Example 52 The compound obtained in Example 1 was treated in the same manner as in Example 13 by using trifluoroacetic acid instead of the hydrochloric acid-dioxane solution, to give amorphous (3S) -2-L-tryptofil- 1,2,3,4-tetrahydroisoquinoline-3-
A carboxylic acid / benzyl ester / trifluoroacetic acid salt was obtained. IR (KBr, cm ^-1 ): 3420, 3035, 173
0, 1675, 1460, 1205, 745 MS (SIMS): 454 (M + 1) Example 53 2.3 g of the compound obtained in Example 7 and 607 mg of N-methylmorpholine were dissolved in 20 ml of tetrahydrofuran, and the solution was dissolved at -15 ° C. Butoxycarbonyl chloride 81
After 9 mg was added dropwise, the mixture was stirred at the same temperature for 10 minutes. After 20 ml of a saturated ammonia / tetrahydrofuran solution was added dropwise, and the mixture was stirred at -15 ° C for 1 hour, the reaction solution was concentrated under reduced pressure. The residue was extracted with chloroform, and the extract was washed, dried and concentrated. The residue was purified by silica gel chromatography (chloroform: methanol = 20: 1) to obtain amorphous (3S) -2- (N-tert- Butyloxycarbonyl-L-tryptofile) -1,2,2
1.62 g of 3,4-tetrahydroisoquinoline-3-carboxamide was obtained. IR (KBr, cm ^-1 ): 3340, 2980, 170
0, 1675, 1630, 1440, 1170, 740 MS (SIMS): 463 (M + 1) Example 54 1.6 g of the compound obtained in Example 53 was treated in the same manner as in Example 13 to give a pale red powder. 1.3 g of (3S) -2- (L-tryptofile) -1,2,3,4-tetrahydroisoquinoline-3-carboxamide hydrochloride was obtained. IR (KBr, cm ^-1 ): 3425, 2900, 167
5,1630,1480,1460,1360,112
0,745 MS (SIMS): 363 (M + 1) Example 55 The compound obtained in Example 53 was treated in the same manner as in Example 13 using trifluoroacetic acid instead of the hydrochloric acid-dioxane solution to give a pale red powder. (3S) -2- (L-tryptofile) -1,2,3,4-tetrahydroisoquinoline-3-carboxamide trifluoroacetate in the form of MS (SIMS): 363 (M + 1) Example 56 (1) 0.5% glucose (manufactured by Nakarai Co., Ltd.), 2
% Soya flower A (manufactured by Nisshin Oil Co., Ltd.), 2% glycerol (manufactured by Nakarai), 0.2% yeast extract (manufactured by Asahi Breweries), 0.25% sodium chloride (manufactured by Nakarai), A liquid medium (pH 7.0) composed of 0.4% calcium carbonate (manufactured by Nitto Powder Chemical Co., Ltd.)
0) Aspergi cultured on agar slope in 100 ml
lulus sp. A374 strain (FERM BP-611)
3) was inoculated with a platinum loop and cultured with shaking at 27 ° C. for 3 days.

The obtained culture was used as a seed culture. One hundred ml of a seed culture solution was inoculated into 100 Erlenmeyer flasks containing 100 ml of the liquid medium having the above composition, and cultured at 27 ° C. for 5 days with shaking.

(2) The bacterial cells were removed by filtration from 85 L of the culture solution obtained by the method (1), and the culture solution was diluted with 3 L of Diaion HP-20 (manufactured by Mitsubishi Chemical Corporation).
After passing through the column, TMC-2A, TMC-2B and TMC-2C were adsorbed on the column. 5 L, 30 L, 75 L, and 30 L of distilled water, 20%, 50%, and 100% methanol respectively were allowed to flow down the column. A 50% methanol fraction exhibiting dipeptidyl peptidase IV inhibitory activity was concentrated under reduced pressure to obtain a crude substance. This crude substance was converted to silica gel (Wakogel C-300, manufactured by Wako Pure Chemical Industries, Ltd., 6
(0 × 900 mm). The elution was performed by first mixing 5 L of a dichloromethane: methanol: ethanol (10: 4: 4) mixed solution, and then increasing the ratio of water as follows in a mixed solution of dichloromethane: methanol: ethanol: water. That is, a mixed solution of dichloromethane: methanol: ethanol: water is mixed with 1
5 L of 0: 4: 4: 0.1 solution, 5 L of 10: 4: 4: 0.2 solution, 10 L of 10: 4: 4: 0.5 solution, 10: 4:
10 L of 4: 1 solution, and finally 10 L of 10: 4: 4: 2 solution
Eluted. Fractions showing dipeptidyl peptidase IV inhibitory activity were collected and concentrated under reduced pressure to obtain a crude substance.

This crude substance was subjected to a reverse phase silica gel column (OD
SA60, manufactured by YMC Co., Ltd., 60 × 900 m
Chromatography was performed using m). Elution is
Performed using 20% acetonitrile-80% water. Each eluted fraction is analyzed by high performance liquid chromatography, and TMC
Fractions containing only -2A were collected, concentrated under reduced pressure, and lyophilized. Analytical high performance liquid chromatography was performed using YMC-Pa
ck AM-301-3 (manufactured by YMC Corporation)
Using a 4.6 × 100 mm column, acetonitrile 10
% To 35% with a 15 minute linear gradient (flow 1.2 ml / min). The detection was based on the absorbance at 210 nm and 254 nm. By the above operation, about 1.6 g of pure TMC-2A could be obtained.

Purification of TMC-2B and TMC-2C was performed as follows. Fractions containing TMC-2B and TMC-2C were collected in the above reverse phase chromatography and concentrated under reduced pressure. The concentrate is subjected to silica gel chromatography (Wakogel C-300, Wako Pure Chemical Industries, Ltd., 22 × 500 m
m) was used to fractionate TMC-2B and TMC-2C.
Elution was performed using dichloromethane: methanol: ethanol (1
0: 4: 4) After flowing 500 ml of the mixed solution, 200 ml of a 10: 4: 4: 0.1 solution of a dichloromethane: methanol: ethanol: water solution was used, and 10: 4: 4:
200 ml of 0.2 solution, 10: 4: 4: 0.5 solution 20
0 ml, 500 ml of a 10: 4: 4: 1 solution and 10:
500 ml of a 4: 4: 2 solution was allowed to flow down sequentially. Each eluted fraction was subjected to silica gel TLC (silica gel; Merck Co. No.
5715, developing solvent; dichloromethane: methanol: ethanol: water = 10: 4: 4: 2).
Fractions containing only TMC-2B or TMC-2C were concentrated to dryness, respectively. By the above operation, pure TMC
5.4 mg and 21 mg of -2B and TMC-2C were obtained, respectively.

[0160]

EFFECT OF THE INVENTION The tetrahydroisoquinoline derivative of the present invention selectively inhibits dipeptidyl peptidase IV at a low concentration, and prevents and treats autoimmune diseases (immune disorders and immunodeficiencies) such as arthritis and rheumatoid arthritis. Useful as an agent.

[Brief description of the drawings]

FIG. 1 is a UV spectrum of TMC-2A.

FIG. 2 is a UV spectrum of TMC-2B.

FIG. 3 is a UV spectrum of TMC-2C.

FIG. 4 is an IR spectrum of TMC-2A.

FIG. 5 is an IR spectrum of TMC-2B.

FIG. 6 is an IR spectrum of TMC-2C.

FIG. 7 is a ¹ H-NMR spectrum of TMC-2A.

FIG. 8 is a ¹ H-NMR spectrum of TMC-2B.

FIG. 9 is a ¹ H-NMR spectrum of TMC-2C.

FIG. 10 is a ¹³ C-NMR spectrum of TMC-2A.

FIG. 11 is a ¹³ C-NMR spectrum of TMC-2B.

FIG. 12 is a ¹³ C-NMR spectrum of TMC-2C.

FIG. 13 is a graph showing the inhibitory effect of TMC-2A on rat alkyldiamine-induced arthritis. Cont (−): control group not receiving alkyldiamine Cont (+): control group receiving alkyldiamine

FIG. 14 is a graph showing the inhibitory effect of TMC-2A and the compound of Example 13 on rat adjuvant-induced arthritis. Cont (-): control group not administered with heat-killed M. tuberculosis Cont (+): control group administered with heat-killed M. tuberculosis

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification code FI C07K 5/08 C12N 9/99 C12N 9/99 C12P 21/02 A C12P 21/02 A61K 37/02 // (C12P 21/02 C72R 1:66) (72) Inventor Nobuaki Nonaka 4-39-10 Sasame, Toda City, Saitama Prefecture Green Heights 202 (72) Inventor Yasuyuki Asai 2250 Sanmuro, Urawa-shi, Saitama Room 103, Yayoi Grand Palace

Claims (25)

[Claims] 1. A pharmaceutical composition comprising a compound having a dipeptidyl peptidase IV inhibitory activity and having a tetrahydroisoquinoline skeleton or a pharmaceutically acceptable salt thereof as an active ingredient. 2. It has a dipeptidyl peptidase IV inhibitory activity and has the formula: A pharmaceutical composition comprising, as an active ingredient, a compound having a partial structure represented by: or a pharmaceutically acceptable salt thereof. 3. A compound of the general formula [I]: (Wherein, R ¹ is (1) a group having a structure in which a hydroxy group of a carboxy group is removed from an amino acid whose amino group may be protected, or (2) a protecting group of an amino group, and R ² is ( 1) an optionally protected hydroxyl group, (2)
A group having a structure in which one hydrogen atom of an amino group is removed from an amino acid whose carboxy group may be protected, or (3) a hydrogen atom on a nitrogen atom is removed from a primary or secondary amine or ammonia. Wherein R ³ , R ⁴ , R ⁵ and R ⁶ are the same or different and each represent a hydrogen atom, a hydroxyl group or a lower alkoxy group) or a pharmaceutically acceptable salt thereof. A pharmaceutical composition as an ingredient. 4. A hydroxy group of a carboxy group is removed from an amino acid in which R ¹ is (1) an amino group whose amino group may be protected by an aryloxycarbonyl group, an aryl-substituted lower alkoxycarbonyl group or a lower alkoxycarbonyl group. Or a group having a modified structure, or (2) an aryloxycarbonyl group, an aryl-substituted lower alkoxycarbonyl group or a lower alkoxycarbonyl group,
R ² represents (1) a hydroxyl group optionally substituted with an aryl-substituted lower alkyl group, or (2) a hydrogen atom of an amino group from an amino acid whose carboxy group may be protected with a lower alkyl or an aryl-substituted lower alkyl. One atom
A group having a removed structure, or (3) an amino group optionally substituted with one or two groups selected from lower alkyl or aryl-substituted lower alkyl, R ³ , R ⁴ , R
The pharmaceutical composition according to claim 3, wherein ⁵ and R ⁶ are the same or different and are a hydrogen atom, a hydroxyl group or a lower alkoxy group. Wherein R ¹ is (1) a benzyloxycarbonyl group or a tert- butoxycarbonyl group with an amino atomic group which may be protected by tryptophyl group, lysyl group or phenylalanyl group or (2) a benzyloxycarbonyl group or tert- butoxycarbonyl group, R ² is (1) hydroxyl group which may be protected with a benzyl group, (2) carboxy atomic methyl group or benzyl group may be protected, alanine, valine, leucine, Isoleucine, proline, phenylalanine,
Tryptophan, methionine, glycine, serine, O-
Benzyl-serine, threonine, cysteine, glutamine, asparagine, tyrosine, lysine, arginine, histidine, aspartic acid, glutamic acid and the formula: Wherein one of R ⁷ and R ⁸ represents a hydroxyl group and the other represents a hydrogen atom or a hydroxyl group, the group having a structure in which one hydrogen atom of the α-amino atom group of the α-amino acid is removed or (3 R) an amino group optionally substituted with one or two groups selected from a tert-butyl group or a benzyl group,
The pharmaceutical composition according to claim 4, wherein ³ is a hydrogen atom or a lower alkoxy group, R ⁴ is a hydrogen atom or a hydroxyl group, R ⁵ is a hydrogen atom or a lower alkoxy group, and R ⁶ is a hydrogen atom, a hydroxyl group or a lower alkoxy group. Wherein R ¹ is tryptophyl group, R ² is (1) hydroxyl group, (2) alanine, valine, leucine, isoleucine, proline, phenylalanine, tryptophan, methionine, glycine, serine, threonine, cysteine, glutamine, asparagine, A group having a structure in which one hydrogen atom of an α-amino atom group has been removed from an α-amino acid selected from tyrosine, lysine, arginine, histidine, aspartic acid and glutamic acid, or (3) an amino group, R ³ , R ⁴ , R ^{5. A method according to} claim 5, wherein ⁵ and R ⁶ are hydrogen atoms.
The pharmaceutical composition according to any one of the preceding claims. 7. R ¹ is tryptophyl group, R ² is (1) hydroxyl group, (2) glutamine, serine, aspartic acid, glutamic acid, alanine, cysteine, arginine, alpha-amino from alpha-amino acid selected from methionine and asparagine 7. The pharmaceutical composition according to claim ⁶ , wherein the group having a structure obtained by removing one hydrogen atom from the atomic group or (3) the amino group, R ³ , R ⁴ , R ⁵ and R ⁶ is a hydrogen atom. 8. A group having a structure in which R ¹ is a tryptophyl group, R ² is (1) a hydroxyl group, (2) an α-amino acid selected from glutamine and serine and one hydrogen atom of an α-amino atomic group is removed or (3) amino group, R ³ , R ⁴ , R ⁵
The pharmaceutical composition according to claim 7, wherein R ⁶ and R ⁶ are a hydrogen atom. 9. A 2-L-tryptofil-1,2,3,9.
A pharmaceutical composition comprising 4-tetrahydroisoquinolyl-3-carboxylic acid and a pharmaceutically acceptable salt thereof as an active ingredient. 10. The general formula [II]: (Wherein one of R ⁷ and R ⁸ represents a hydroxyl group and the other represents a hydrogen atom or a hydroxyl group) or a pharmacologically acceptable salt thereof as an active ingredient. 11. The pharmaceutical composition according to claim 3, wherein the pharmaceutical composition is a dipeptidyl peptidase IV inhibitor. 12. The pharmaceutical composition according to claim 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, which is a prophylactic / therapeutic agent for an autoimmune disease. 13. The pharmaceutical composition according to claim 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 which is an agent for preventing or treating arthritis. 14. The agent according to claim 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 which is a prophylactic / therapeutic agent for rheumatoid arthritis.
The pharmaceutical composition according to any one of the preceding claims. 15. A compound of the general formula [I]: (Wherein, R ¹ is (1) a group having a structure in which a hydroxy group of a carboxy group is removed from an amino acid whose amino group may be protected, or (2) a protecting group of an amino group, and R ² is ( 1) an optionally protected hydroxyl group, (2)
A group having a structure in which one hydrogen atom of an amino group is removed from an amino acid whose carboxy group may be protected, or (3) a hydrogen atom on a nitrogen atom is removed from a primary or secondary amine or ammonia. Wherein R ³ , R ⁴ , R ⁵ and R ⁶ are the same or different and each represent a hydrogen atom, a hydroxyl group or a lower alkoxy group) or a pharmaceutically acceptable salt thereof. 16. A hydroxy group of a carboxy group is removed from an amino acid wherein R ¹ is (1) an amino group whose amino group may be protected by an aryloxycarbonyl group, an aryl-substituted lower alkoxycarbonyl group or a lower alkoxycarbonyl group. A group having a substituted structure, (2) an aryloxycarbonyl group, an aryl-substituted lower alkoxycarbonyl group or a lower alkoxycarbonyl group, a hydroxyl group in which R ² may be substituted with (1) an aryl-substituted lower alkyl group, (2 A) a group having a structure in which one hydrogen atom of an amino group has been removed from an amino acid in which a carboxyl group may be protected by a lower alkyl or aryl group-substituted lower alkyl; or (3) a lower alkyl or aryl-substituted lower alkyl Substituted with one or two groups selected from It is an amino group, R ^3,
16. The tetrahydroisoquinoline derivative or a pharmaceutically acceptable salt thereof according to claim 15, wherein R ⁴ , R ⁵ and R ⁶ are the same or different and are a hydrogen atom, a hydroxyl group or a lower alkoxy group. 17. R ¹ is (1) a benzyloxycarbonyl group or a tert- butoxycarbonyl group with an amino atomic group may tryptophyl group which may be protected, lysyl group or phenylalanyl group or (2) a benzyloxycarbonyl group Or tert-butoxycarbonyl group, R ² is (1) a hydroxyl group optionally protected with a benzyl group, (2) a carboxy atom group optionally protected with a methyl group or a benzyl group, alanine, valine,
Leucine, isoleucine, proline, phenylalanine, tryptophan, methionine, glycine, serine,
O-benzyl-serine, threonine, cysteine, glutamine, asparagine, tyrosine, lysine, arginine, histidine, aspartic acid, glutamic acid and the formula: Wherein one of R ⁷ and R ⁸ represents a hydroxyl group and the other represents a hydrogen atom or a hydroxyl group, the group having a structure in which one hydrogen atom of the α-amino atom group of the α-amino acid is removed or (3 R) an amino group optionally substituted with one or two groups selected from a tert-butyl group or a benzyl group,
³ is a hydrogen atom or a lower alkoxy group, R ⁴ is a hydrogen atom or a hydroxyl group, R ⁵ is a hydrogen atom or a lower alkoxy group, and R ⁶ is a hydrogen atom, a hydroxyl group or a lower alkoxy group.
Or a pharmacologically acceptable salt thereof. 18. R ¹ is a tryptophyll group and R ² is (1)
Hydroxyl group, (2) alanine, valine, leucine, isoleucine, proline, phenylalanine, tryptophan,
A group having a structure in which one hydrogen atom of an α-amino group has been removed from an α-amino acid selected from methionine, glycine, serine, threonine, cysteine, glutamine, asparagine, tyrosine, lysine, arginine, histidine, aspartic acid and glutamic acid. Or (3) the amino group, R ³ , R ⁴ , R ⁵ and R ⁶ are hydrogen atoms.
7. The tetrahydroisoquinoline derivative according to 7, or a pharmaceutically acceptable salt thereof. 19. R ¹ is a tryptophyll group and R ² is (1)
Hydroxyl group, (2) glutamine, serine, aspartic acid,
A group having a structure in which one hydrogen atom of an α-amino group has been removed from an α-amino acid selected from glutamic acid, alanine, cysteine, arginine, methionine and asparagine, or (3) an amino group, R ³ , R ⁴ , R ⁵ 19. The tetrahydroisoquinoline derivative according to claim 18, wherein R ⁶ is a hydrogen atom, or a pharmaceutically acceptable salt thereof. 20. R ¹ is a tryptophyll group and R ² is (1)
Hydroxyl group, (2) α- selected from glutamine and serine
A group having a structure in which one hydrogen atom of an α-amino group has been removed from an amino acid, or (3) an amino group, R ³ , R ⁴ , R
20. The tetrahydroisoquinoline derivative or a pharmaceutically acceptable salt thereof according to claim 19, wherein ⁵ and R ⁶ are hydrogen atoms. 21. 2-L-tryptofil-1,2,2
3,4-tetrahydroisoquinolyl-3-carboxylic acid and pharmaceutically acceptable salts thereof. 22. The general formula [II]: Wherein one of R ⁷ and R ⁸ represents a hydroxyl group and the other represents a hydrogen atom or a hydroxyl group, or a pharmaceutically acceptable salt thereof. 23. The general formula [II]: Wherein one of R ⁷ and R ⁸ represents a hydroxyl group and the other represents a hydrogen atom or a hydroxyl group, and a carbon source and nitrogen capable of assimilating mold belonging to the genus Aspergillus. A method for producing a tetrahydroisoquinoline derivative, comprising culturing in a nutrient medium containing a source, and isolating compound [II] from the culture. 24. The general formula [III]: (Wherein, R ¹¹ represents (1) a group having a structure in which at least an amino group in which an amino group is protected is obtained by removing a hydroxy group of a carboxy group, or (2) a protecting group of an amino group, R ³ and R ⁴ , R ⁵ and R ⁶ are the same or different and each represents a hydrogen atom, a hydroxyl group or a lower alkoxy group) or a reactive derivative of the carboxy group thereof and a general formula [IV]: (Wherein R ²¹ represents (1) a hydrogen atom of an amino group from an amino acid having at least a protected carboxy group)
A group having a removed structure or (2) primary or secondary
Hydrogen atom on nitrogen atom from primary amine or ammonia
(Representing a group having a removed structure) by condensation with a compound represented by the general formula [V]: (Wherein R ¹¹ , R ²¹ , R ³ , R ⁴ , R ⁵ and R ⁶ have the same meanings as described above), and if necessary, the protecting group is removed. Formula [Ia]: (Wherein, R ¹ is (1) a group having a structure in which a hydroxy group of a carboxy group is removed from an amino acid whose amino group may be protected or (2) a protecting group of an amino group, and R ²² is ( 1) a group having a structure in which one hydrogen atom of an amino atom group has been removed from an amino acid whose carboxy atom group may be protected, or (2) one hydrogen atom on a nitrogen atom from a primary or secondary amine or ammonia. And R ³ , R ⁴ , R ⁵ and R ⁶ have the same meaning as described above). 25. The general formula [VI]: (Wherein R ²³ is (1) a protected hydroxyl group, (2) a group having a structure in which at least one hydrogen atom of an amino group has been removed from an amino acid having at least a protected carboxy group, or (3) 1 A group having a structure in which one hydrogen atom on a nitrogen atom has been removed from a secondary or secondary amine or ammonia, and R ³ , R ⁴ , R ⁵ and R ⁶ are the same or different and represent a hydrogen atom, a hydroxyl group or a lower alkoxy group ) And a compound of the general formula [VII]: (Wherein, R ¹² represents a group having a structure in which at least the amino group is protected and the hydroxy group of the carboxy group is removed from the protected amino group) or a reactive derivative of the carboxy group thereof. To give a compound of the general formula [VIII]: Wherein R ¹² , R ²³ , R ³ , R ⁴ , R ⁵ and R ⁶ have the same meanings as described above, and if necessary, the protecting group is removed. Formula [Ib]: (Wherein, R ¹³ is a group having a structure in which a hydroxy group of a carboxy group is removed from an amino acid whose amino group may be substituted, R ² is (1) a hydroxyl group which may be protected, (2 A) a group having a structure in which one hydrogen atom of an amino atom group has been removed from an amino acid whose carboxy atom group may be protected, or (3) one hydrogen atom on a nitrogen atom from a primary or secondary amine or ammonia. Wherein R ³ , R ⁴ , R ⁵ and R ⁶ have the same meaning as described above).